Anther culture in apple (Malus domestica Borkh), improve breeding program. Anther at the mid uninucleate stage of pollen development were obtained from two apple genotypes cultivars and stored for one week in refrigerator at 4 ºC. Anther was cultured on MS medium supplemented with 5% sucrose and combination of phytohormones [6-Benzylaminopurine (BA), thidiazuron (TDZ), indol-3-butyric acid (IBA), Naphthalene acetic acid (NAA)] for callus induction and regeneration. Addition of TDZ+IBA to multiplication medium were more effective for increasing the number of initiated shoots compared with BA, NAA. The average percentage of callus formation from Dorsett golden cultivar 31.64% in the first season, while it was 28.38% in the second season. Anna revealed a 27.86% & 25.99% of callus formation in the first &second season, respectively. Regeneration percentage was 24.86%& 23.82% with Dorsett golden, however, it was 20.57% &19.79% with Anna in the first and second seasons, respectively. Positive correlation between media and genotype was showed in first season only for plant regeneration. Moreover, a negative correlation was revealed between media and genotype for callus induction in both seasons. Cytological studies revealed normal cells ranged from 96.90%-97.85% and abnormal cells of about 2.15%-3.01%.